1 鄂州市中心医院内分泌科，湖北 鄂州 436000
2 华中科技大学同济医学院附属梨园医院中西医结合科，武汉 430077
3 武汉长江航运总医院肾内科，武汉 430010
|| 国家自然科学基金 (81673657)。
目的：观察贞清方对糖尿病肾病(diabetic nephropathy，DN)大鼠凝血纤溶系统的调节作用，并探讨其作用与剂量的关系。方法：将Wistar大鼠随机分为正常组、模型组、贞清方高剂量组和贞清方低剂量组。除正常组外，均用腹腔注射链脲佐菌素(streptozotocin，STZ)的方法建立糖尿病模型。检测喂养1个月及注射STZ后1周各组大鼠的血胰岛素(fasting ser um lisulin，FINS)水平，喂养1个月、注射STZ后1周、给药4周及给药8周后的血糖(fasting blood glucose，FBG)、血脂(TC、 TG)水平、给药8周后的各组大鼠血浆血栓素B2(thromboxane B2，TXB2)、组织型纤溶酶原激活物(tissue plasminogen activator，t-PA)和纤溶酶原激活物抑制物-1(plasminogen activator inhibitor type-1，PAI-1)含量及各组大鼠尿微量白蛋白(UAE)、血肌酐(SCr)和尿肌酐(UCr)的改变，并计算内生肌酐清除率(CCr)。称量大鼠体重后处死，称量肾重，并计算肾重指数(kidney index，KI)、将肾组织进行HE染色、过碘酸希夫(PAS)染色观察肾病变，并用免疫组织化学及Western印迹检测肾组织PAI-1的蛋白表达水平及用聚合酶链反应(PCR)检测肾皮质PAI-1的mRNA的表达变化。结果：模型组大鼠各时期的FIN，FBG，TC，TG均显著高于正常组(P<0.05)，与正常组相比，模型组血浆KI，TXB2，PAI-1，UAE，CCr也明显升高(P<0.05)，肾组织PAI-1蛋白表达及mRNA水平明显上调(P<0.05)，而治疗组以上指标均低于模型组(P<0.05)，且贞清方高剂量组低于贞清方低剂量组。 结论：贞清方可明显降低血糖，血脂；升高t-PA的水平，降低血浆PAI-1，TXB2含量，减少糖尿病大鼠肾组织PAI-1的蛋白和mRNA的表达量，且贞清方对糖尿病大鼠肾的保护作用呈剂量依赖性。
Protective effect of Zhenqing Recipe on kidney of diabetic nephropathy rats at different doses
CorrespondingAuthor: WEN Xiuying
This work was supported by the National Natural Science Foundation of China (81673657).
Objective: To observe the protective effect of Zhenqing Recipe on coagulation and fibrinolysis system in rats with diabetic nephropathy, and to explore the relationship between effects and doses. Methods: The Wistar rats were randomly divided into a normal group, a model group, a high dose group of Zhenqing Recipe and the low dose of Zhenqing Recipe group. Diabetic models were established by injecting streptozotocin (STZ) into the abdomen except for the normal group. The blood insulin (FINS) of each group’s rats after feeding 1 month and 1 week after the injecting STZ were detected. The blood glucose (FBG), blood lipid (TC, TG) level after feeding 1 month, 1 week after the injection of STZ, and 4 or 8 weeks after feeding decoction, the plasma thromboxane B2 (TXB2), tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) content and urine trace albumin (UAE), serum creatinine (SCr) and the changes of urine creatinine (UCr) of each rat after dosing 8 weeks were detected. The endogenous creatinine clearance rate (CCr) was calculated. These rats were weighed and then put to death. Calculated the kidney weight index (KI). Hematoxylin staining (HE) and periodic acid Schiff staining were used to observe renal lesions. The immunohistochemistry and protein immunoblotting (Western blot) were used to test expression levels of PAI-1, and the polymerase chain reaction was used to decect PAI-1 mRNA expression of renal tissues. Results: The FIN, FBG, TC, TG of pathological group rats in each period were significantly higher than that of normal control group (P<0.05). Compared with normal group, the plasma KI, TXB2, PAI-1, UAE, and CCr of the pathological group were also increased significantly (P<0.05), and the expressions of proteins and mRNA levels of PAI-1significantly raised of renal tissues (P<0.05), while these indicators of treatment group are lower than the model group (P<0.05), which in high dose group were lower than the low dose group. Conclusion: Zhenqing Recipe can obviously reduce blood glucose, blood lipid levels, increase the levels of t-PA, reduce plasma PAI-1, TXB2 levels, reduce the expressions of protein and mRNA of kidney PAI-1 of diabetic rats. The renal protective effect on diabetic rats of Zhenqing Recipe and was dose-dependent.
Zhenqing Recipe; diabetic nephropathy; tissue type plasminogen activator; plasminogen activator inhibitor-1; thromboxane B2